Root-lesion nematodes, Pratylenchus sp., are recognized as one of the most important plant parasitic nematodes. They infest various crop plants and cause severe root damages which lead to the yield loss of agricultural crops. There are 72 species recorded in the world, but 14 species in Korea. Soil samples were collected from chrysanthemum cultivars in various regions. Morphological and molecular diagnosis firstly recognized two species as Pratylenchus kumamotoensis and P. pseudocoffeae in Korea. Morphologically, P. kumamotoensis characterized by pharyngeal gland lobes overlapping with intestine dorsally. P. pseudocoffeae, which is similar with P. coffeae, but pharyngeal gland lobes are longer than P. coffeae. In molecular diagnosis, nucleotide sequences of D2-D3 region (700-800 bp) within 28S rDNA and ITS region including ITS1, 5.8S and ITS2 (700-1,000 bp) between 18S and 28S rDNA were determined from collected samples. NCBI database comparison of D2-D3 of P. kumamotoensis and P. pseudocoffeae were 99% similar with those of corresponding species, respectively. ITS of P. pseudocoffeae was 99% similar within species. ITS sequence of P. kumamotoensis was firstly determined in this study. Distribution of P. kumamotoensis was only reported in Japan but P. pseudocoffeae is present in Japan and USA. Here we firstly report the presence of these two species in Korea.